Mast cells are critical mediators in the initiation and propagation of allergic inflammation by secreting histamine and pro-inflammatory cytokines. Current treatments, including corticosteroids and antihistamines, provide symptomatic relief but do not directly target FcεRI-mediated mast cell activation. Esculetin, a natural bioactive compound from Fraxinus rhynchophylla Hance with potent antioxidative and anti-inflammatory effects, offers a promising but underexplored strategy for modulating allergic responses. This study investigates the pharmacological potential of esculetin in suppressing mast cell–mediated allergic inflammation. Esculetin was orally administered in immunoglobulin (Ig)E-mediated passive cutaneous anaphylaxis (PCA) and ovalbumin-induced active systemic anaphylaxis (ASA) mouse models. PCA and ASA responses were assessed by dye extravasation, ear swelling, hypothermia, and serum levels of IgE, IL-4, and histamine. In vitro studies using RBL-2H3 cells and bone marrow–derived mast cells examined FcεRI signaling and degranulation. Esculetin administered orally led to a dose-dependent inhibition of PCA responses, reflected by decreased Evans blue dye extravasation, reduced ear tissue swelling, and suppressed mast cell degranulation. In the ASA model, esculetin markedly alleviated hypothermia and significantly reduced serum levels of IgE, interleukin (IL)-4, and histamine. In vitro analyses using RBL-2H3 cells and murine bone marrow–derived mast cells showed that esculetin suppressed degranulation by attenuating FcεRI signaling cascades. Specifically, esculetin targeted Src family kinases such as Lyn and Fyn, as well as the downstream kinase Syk, ultimately reducing intracellular calcium influx. These findings suggest that esculetin may serve as a promising therapeutic candidate for mast cell-mediated allergic inflammatory disorders by inhibiting FcεRI signaling, thereby suppressing mast cell activation.

The interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) signaling pathway plays a pivotal role in regulating inflammation and tumorigenesis, making it a significant target for therapeutic intervention. Although the rhizome of Curcuma longa (C. longa) has been extensively investigated, the pharmacological potential of its aerial parts, such as the leaves, remains largely unexplored. The present study investigated the inhibitory effects of the ethyl acetate fraction of C. longa leaf extract (CL-E) on IL-6-induced STAT3 activation in Hep3B hepatoma cells in vitro. CL-E significantly suppressed IL-6-stimulated STAT3 transcriptional activity in a dose-dependent manner (IC50, 33.9±3.7 µg/ml) without compromising cell viability (up to 60 µg/ml). Mechanistically, CL-E reduced STAT3 phosphorylation at Tyr705 while enhancing STAT3 phosphorylation at Ser727; this opposing pattern suggests an ERK-mediated rebalancing of STAT3 signaling that attenuates canonical Tyr705-dependent activation and augments Ser727-associated modulation. Additionally, CL-E inhibited STAT3 nuclear translocation and downregulated the mRNA expression of STAT3 target genes, including C-reactive protein and suppressor of cytokine signaling 3. Mechanistic investigations revealed that CL-E promoted extracellular signal-regulated kinase (ERK)1/2 phosphorylation, leading to increased STAT3 Ser727 phosphorylation. Notably, the inhibitory effect of CL-E on STAT3 Tyr705 phosphorylation was reversed by MEK1/2 (U0126) or PKC inhibitors (bisindolylmaleimide II), indicating that CL-E modulates STAT3 signaling through an ERK-mediated mechanism. Collectively, these in vitro findings identify C. longa leaves as an underutilized botanical resource with the potential to regulate IL-6/STAT3 signaling, warranting in vivo evaluation to establish its efficacy, safety and pharmacokinetics, and to define potential therapeutic utility in IL-6/STAT3-driven conditions.

Undaria pinnatifida ( U. pinnatifida) has long been a part of the human diet and medicine. Although U. pinnatifida has been reported to have immunomodulatory, anti-inflammatory, anti-diabetic and antibacterial activities, its specific effect on patients with combined allergic rhinitis and asthma syndrome (CARAS) has not been clarified. In this study, the anti-allergic and anti-inflammatory effects of U. pinnatifida extract (UPE) are investigated in a mouse model of ovalbumin (OVA)-induced CARAS. The oral administration of UPE inhibits allergic responses by reducing OVA-specific immunoglobulin levels. As a result, the symptoms of early reactions are also improved. UPE inhibits the accumulation of inflammatory cells and attenuates the expression of Th2 cytokines in both nasal and bronchoalveolar lavage fluid. Furthermore, UPE treatment inhibits the NF-κB/MAPK signaling pathway in lung homogenates. Additionally, UPE prevents shedding of the nasal mucosal epithelium, protects the integrity of the epithelium, enhances the expression of E-cadherin at the junction of epithelial cells, and inhibits the degradation of ZO-1 and occludin in the airway epithelium. In addition, UPE ameliorates dysfunction of the nasal epithelial barrier by enhancing antioxidant properties and downregulating the expression of the inflammatory factor IL-33. These results suggest that UPE may treat CARAS by modulating epithelial cell dysfunction and oxidative stress.

Protectin D1 (PD1) derived from docosahexaenoic acid (DHA) has shown promise in resolving inflammation. Mast cells are critical drivers of allergic inflammation, releasing inflammatory mediators such as histamine and pro-inflammatory cytokines. This study assesses the effectiveness of PD1 in counteracting mast cell-mediated allergic inflammation. In vivo, two well-established mouse models were employed: IgE-mediated passive cutaneous anaphylaxis (PCA) and ovalbumin-induced active systemic anaphylaxis (ASA). The oral administration of PD1 markedly suppressed PCA reactions, including ear swelling, plasma extravasation of Evans blue and mast cell degranulation. In the ASA model, oral PD1 administration dose-dependently alleviated hypothermia and reduced elevated serum levels of IgE, histamine, and IL-4. Mechanistic insights were gained through studies in the RBL-2H3 and primary mast cells derived from mouse bone marrow, where PD1 inhibited IgE-mediated degranulation and decreased intracellular calcium influx by blocking FcεRⅠ signaling pathways involving Lyn, Fyn, and Syk kinases. Additionally, PD1 suppressed pro-inflammatory cytokine production by inhibiting the activity of critical transcription factor; nuclear factor-κB. These findings suggest that PD1, a bioactive lipid derived from DHA, is a very promising therapeutic candidate for mast cell-derived allergic inflammation.

This study evaluated the immune-enhancing efficacy of Limosilactobacillus fermentum KBL375 isolated from the feces of healthy Koreans. KBL375-treated splenocytes showed enhancement of cytotoxicity against YAC-1 cells, the target of natural killer (NK) cells, with an increase in CD335, granzyme B, perforin, and interferon-gamma (IFN-γ). Oral administration of KBL375 in mice with cyclophosphamide (CP)-induced immunosuppression improved body weight and immune functions, including immune organ indices, lymphocyte proliferations, and immunoglobulin (Ig) A levels. Notably, KBL375 increased NK cell cytotoxicity and proportion in immunosuppressed mice. Perforin/IFN-γ expression levels, which indicated NK cell activation, were also increased in KBL375-treated mice. Furthermore, KBL375 led to an increase in beneficial microbes, such as Bifidobacterium, in the gut microbiome of immunosuppressed mice, fostering a favorable intestinal microbial environment. These comprehensive results suggest that KBL375 exhibits potent immune regulatory functions and positively influences the gut microbiota, implying its potential as a probiotic agent for immune enhancement.

This study proposes a novel in vitro-in vivo extrapolation (IVIVE) strategy that integrates a patented biomimetic in vitro system with pharmacokinetic modeling to improve the prediction of in vivo drug kinetics. The system employed a single-well plate design with various sizes of mesh inserts to simultaneously assess drug diffusion and cellular metabolism. Drug diffusion patterns were quantified and modeled using a Weibull distribution to establish mathematical relationships between pore size and kinetic parameters. The model was extended to predict the metabolic phase with two drugs, the metabolic conversion of diclofenac into 4-hydroxydiclofenac and metabolic clearance of testosterone in HepaRG cells. The predicted in vivo hepatic clearance values derived from the in vitro model were consistent with those reported in vivo, validating the approach. This integrated strategy enhances IVIVE accuracy and supports the 3 R (replacement, reduction, and refinement) principle by reducing the reliance on animal testing. The findings demonstrate the potential of combining biomimetic systems and mathematical modeling as a reliable platform for pharmacokinetic prediction in drug development.

Topical corticosteroids are widely used to manage chronic inflammatory skin diseases, such as atopic dermatitis (AD). However, their limited skin penetration necessitates high doses and prolonged use, which increases the risk of local and systemic side effects. To address these limitations, we developed a gold nanorod-conjugated dexamethasone (AuNR-DEX) system combined with red light-emitting diode (LED) irradiation to enhance transdermal drug delivery and therapeutic efficacy. AuNR conjugation enhanced the physicochemical properties of DEX, including its hydrodynamic diameter and surface charge, thereby facilitating increased skin penetration. In an AD mouse model, topical AuNR-DEX significantly alleviated disease symptoms at only 20% of the dose required for free DEX. While AuNR-DEX alone showed limited dispersion across the skin, cotreatment with red LED irradiation facilitated broader spreading and improved drug distribution. The combination therapy markedly restored epidermal differentiation, collagen fiber integrity, and calcium homeostasis in AD skin lesions. In vitro, AuNR-DEX plus LED treatment enhanced keratinocyte survival, restored the localization of tight junction proteins, and reduced mitochondrial calcium overload and reactive oxygen species levels. Overall, the synergistic effect of AuNR-DEX and red LED irradiation enhanced drug delivery, targeted dispersion, and therapeutic outcomes, offering a promising strategy for treating AD with reduced corticosteroid dosage and minimized side effects.

OBJECTIVE:Atopic dermatitis (AD) is a chronic skin disease marked by immune dysregulation such as upregulated T helper (Th) 2 responses. While Th2-targeted therapies for AD are under development, their application is limited by side effects such as hypereosinophilia and arthritis. Amygdalin is a glucoside known for its anti-inflammatory and antioxidant effects. It is an essential component of bitter apricot kernel, traditionally utilized to alleviate inflammatory skin diseases such as boils and acne. This study focused on investigating the therapeutic effects of amygdalin on AD. MATERIALS AND METHODS:Its effectiveness was evaluated both in vivo, using the AD mouse model induced by 2,4-dinitrochlorobenzene (DNCB) and Dermatophagoides farinae extract (DFE), and in vitro, using activated leukemia T lymphoblasts and keratinocytes. RESULTS:Amygdalin was shown to reduce the infiltration of immune cells in lesions and both total and DFE-specific immunoglobulin E (IgE) levels in mouse serum. Of note, it explicitly suppressed the expression of Th2 cytokines including interleukin (IL)-4, IL-5, and IL-13, as well as tumor necrosis factor (TNF)-α in ear tissues with AD induced by DNCB/DFE. These phenomena were corroborated by observations in CCRF-CEM cells, where amygdalin notably reduced the levels of IL-4 and TNF-α by inhibiting nuclear translocation of nuclear factor of activated T cells 1 and nuclear factor-κB. CONCLUSION:These findings suggest that amygdalin effectively alleviates allergic skin inflammation by suppressing the Th2 and inflammatory responses, making it a promising candidate for AD treatment.

Excessive exercise can result in a detrimental condition known as overtraining syndrome (OTS), which is characterized by impaired immune function. In this study, we aimed to explore the immunomodulatory properties of YSK-A, a composite herbal substance comprising equal proportions of Boswellia serrata, Commiphora myrrha, and propolis, using a forced swimming (FS)-induced immunocompromised mouse model. Ex vivo experiments showed that YSK-A significantly increased the activity of NK cells against YAC-1 tumor cells compared with its individual components. HPLC analysis for quality control identified KBA, AKBA, furanoeudesma 1,3-diene, cinnamic acid, and galangin as the five primary components of YSK-A. Following sacrifice, improved profiles of molecules related to OTS, immune organ indices, white blood cell proportions in blood, and serum immunoglobulin levels were observed. Furthermore, YSK-A was found to enhance immunocyte function, including cytotoxic effect of splenic NK cells, and proliferation of lymphocytes. Our study suggests that YSK-A exhibits immunomodulatory effects against immune dysfunction.

Nypa fruticans Wurmb is known to contain large amounts of polyphenols and flavonoids with antioxidative and anti-inflammatory effects. However, the biological and physiological functions of N. fruticans have not been scientifically investigated. Thus, we investigated the immunomodulatory effect of N. fruticans hot water extract (YSK-N) in mice using an immune compromised model established by forced swimming (FS). Intensive exercise decreased body weight, organ index, and various immunological parameters in FS mice. However, oral administration of YSK-N significantly restored the FS-induced decreases in body, thymus, and spleen weights, as well as the reduction in the numbers of white blood cells and lymphocytes in the whole blood of mice. Additionally, YSK-N increased splenic cell proliferation in the absence and presence of concanavalin A or lipopolysaccharide stimulation in a concentration-dependent manner. Notably, YSK-N enhanced the cytotoxic activity of natural killer cells against YAC-1 tumor cells under immunosuppressive conditions. Furthermore, YSK-N supplementation reverted the FS-induced downregulation in immunoglobulin production and Il2, Il6, Il12, Ifn gamma, Gzmb, and Prf1 mRNA expression. Therefore, our observations suggested that YSK-N promotes immune function and has potential as an immunomodulatory agent.

Atopic dermatitis (AD), a chronic inflammatory skin disease whose incidence is increasing worldwide, requires the development of alternative treatments due to limited treatment options and concerns about side effects of therapeutic agents. Aspalathin (ASP) is the primary flavonoid found in rooibos, an herb traditionally used for allergies and eczema, accounting for over 40 % of the total flavonoid content, especially in its unfermented state (Green rooibos). This research conducted a thorough investigation into the pharmacological properties of ASP on AD, emphasizing local responses via activated keratinocytes, systemic responses involving T cells and basophils, and an integrated assessment using an AD mouse model. Topical application of ASP significantly reduced AD phenotypes, including erythema, scaling, and increased skin thickness, in AD mouse model. Histological analysis indicated a decrease in the infiltration of immune cells in skin lesions. Moreover, ASP down-regulated inflammatory markers, including T helper (Th)1 and Th2 cytokines, in both skin tissues and activated mouse T cells. In particular, ASP significantly reduced serum immunoglobulin (Ig)E and IgG2a levels. ASP suppressed the expression of cytokines linked to allergy and inflammation in T cells, basophils, and keratinocytes. Mechanistically, ASP exhibited anti-inflammatory properties by inhibiting STAT6 and NFAT1 activation in AD mouse skin and in activated T cells, basophils, and keratinocytes. In conclusion, ASP displayed pronounced effectiveness in relieving AD by sophisticated modulation of immune responses across both local and systemic domains. These findings highlight ASP's promise as a therapeutic intervention for AD, providing a solid scientific basis for future exploration and development.

BACKGROUND:Allergic rhinitis (AR) is the most prevalent form of atopic disease. Undaria pinnatifida has potent antioxidative, antidiabetic, and anti-inflammatory properties.AIMS:We investigated the immunomodulatory effect of Undaria pinnatifida extract (UPE) on allergic inflammation in an AR mouse model.MATERIALS & METHODS:Mice were sensitized and intranasally challenged with ovalbumin (OVA), and the Th1/Th2 and Th17/Treg-related cytokines and histopathology were exanimated after UPE treatments. Enzyme-linked immunosorbent assay was performed using serum samples and NALF to detect OVA-specific immunoglobulins and inflammatory cytokines. Mitogen-activated protein kinases (MAPKs) were measured by western blotting analysis, and an in vitro study measured mast cell activation induced by compound 48/80.RESULTS:After UPE treatment, nasal and lung allergy symptoms, nasal mucosal swelling, and goblet cell hyperplasia were ameliorated. Oral UPE regulated the balance of Th1/Th2 and Th17/Treg cell differentiation in AR mice in a dose-dependent manner. In addition, UPE attenuated the migration of eosinophils and mast cells to the nasal mucosa by suppressing nuclear factor kappa B (NF-κB)/MAPKs. The levels of anti-OVA IgE and IgG1 were also decreased.DISCUSSION:UPE inhibited inflammation by regulating the NF-κB/MAPKs signaling pathway and supressing the activation of critical immune cells such as eosinophils and mast cells.CONCLUSION:UPE may have therapeutic potential for AR.

For centuries, natural products are regarded as vital medicines for human survival. Clematis terniflora var. mandshurica (Rupr.) Ohwi is an ingredient of the herbal medicine, Wei Ling Xian, which has been used in Chinese medicine to alleviate pain, fever, and inflammation. In particular, C. terniflora leaves have been used to cure various inflammatory diseases, including tonsillitis, cholelithiasis, and conjunctivitis. Based on these properties, this study aimed to scientifically investigate the anti-inflammatory effect of an ethanol extract of leaves of C. terniflora (EELCT) using activated macrophages that play central roles in inflammatory response. In this study, EELCT inhibited the essential inflammatory mediators, such as nitric oxide, cyclooxygenase-2, tumor necrosis factor-α, interleukin- (IL-) 6, IL-1β, and inducible nitric oxide synthase, by suppressing the nuclear factor-κB and mitogen-activated protein kinase activation in macrophages. Acute lung injury (ALI) is a fatal respiratory disease accompanied by serious inflammation. With high mortality rate, the disease has no effective treatments. Therefore, new therapeutic agents must be developed for ALI. We expected that EELCT can be a promising therapeutic agent for ALI by reducing inflammatory responses and evaluated its action in a lipopolysaccharide- (LPS-) induced ALI model. EELCT alleviated histological changes, immune cell infiltration, inflammatory mediator production, and protein-rich pulmonary edema during ALI. Collectively, our results may explain the traditional usage of C. terniflora in inflammatory diseases and suggest the promising potential of EELCT as therapeutic candidate for ALI.

Hemodialysis patients are susceptible to cardiovascular remodeling, which increases the risk of cardiovascular morbidity and mortality. Circulating extracellular matrix (ECM)-associated molecules increase during cardiovascular remodeling and can be potential biomarkers of adverse cardiovascular outcomes. However, their clinical significance in patients undergoing hemodialysis remain unclear. This study aimed to elucidate the association between circulating ECM-associated molecules and cardiovascular outcomes in patients undergoing hemodialysis. To this end, we measured levels of plasma matrix metalloproteinase (MMP)-2, MMP-9, tenascin-C, and thrombospondin-2 in 372 patients with hemodialysis. Plasma MMP-2 levels were significantly higher in patients with future cardiovascular events than in those without future cardiovascular events ( P = 0.004). All measured molecules had significant correlations with amino-terminal pro-brain natriuretic peptide levels, but the correlation coefficient was the strongest for plasma MMP-2 (rho = 0.317, P < 0.001). High plasma MMP-2 levels were predictive of left ventricular (LV) diastolic dysfunction (adjusted odds ratio per a standard deviation increase = 1.48, 95% confidence interval [CI] = 1.05–2.08) and were independently associated with an increased risk of composite cardiovascular events (adjusted hazard ratio per a standard deviation increase = 1.30, 95% CI = 1.04–1.63). In conclusion, high plasma MMP-2 levels are associated with LV diastolic dysfunction and an increased risk of adverse cardiovascular outcomes in hemodialysis patients.